Not just a smiley face: Using DNA origami to solve problems in molecular biology

不仅仅是笑脸：利用 DNA 折纸解决分子生物学问题

• 批准号: BB/J018236/1
• 负责人: Robert Henderson
• 金额: $ 78.83万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FJ018236%2F1
• 关键词: just; smiley; face; Using; DNA

The shape of the DNA molecule, a long, string-like double-helical structure is one of the iconic images of modern science. The reason that DNA has this shape is due to the chemicals that make up its structure and which in turn provide an organism's genetic code. A feature of the structure of DNA is that it has recently been shown that it is possible to split the double-helix into two, and long lengths of the resulting 'single-stranded' DNA can be folded into different shapes, producing tiny structures. This technique is called 'DNA origami'. We want to use biotechnology make DNA origami structures, and design them in such a way that we can use them to study two problems that have proved intractable using other experimental methods. We will use a very high resolution 'atomic force microscope' to look at the structures (they are very small - 725 million of them would fit on a 1.5 mm diameter pin-head). The first project is to look at the way that proteins that regulate cells' behaviour find specific points on DNA itself, which they normally identify to turn cell functions on and off. How they do this is puzzling because the proteins are very small and the DNA is very long. The second question concerns how different molecules 'talk' to each other inside a cell in order to produce signals to control of a cell's behaviour. So, we will address this by attaching all of the members of the signalling molecule family to DNA origami tiles in different patterns to show how the proximity of one member to that of another may be influential.

DNA分子的形状，一个长的，像绳子一样的双螺旋结构是现代科学的标志性形象之一。DNA具有这种形状的原因是由于构成其结构的化学物质，这些化学物质反过来又提供了生物体的遗传密码。DNA结构的一个特征是，最近已经表明，双螺旋可以一分为二，并且所产生的“单链”DNA的长长度可以折叠成不同的形状，产生微小的结构。这种技术被称为“DNA折纸”。我们想利用生物技术制造DNA折纸结构，并将其设计成这样一种方式，即我们可以用它们来研究两个问题，这两个问题已经被证明是用其他实验方法难以解决的。我们将使用一个非常高分辨率的“原子力显微镜”来观察这些结构（它们非常小--其中7.25亿个可以装在一个直径为1.5毫米的针头上）。第一个项目是研究调节细胞行为的蛋白质如何在DNA本身上找到特定的点，它们通常会识别这些点来开启和关闭细胞功能。它们是如何做到这一点的令人困惑，因为蛋白质非常小，而DNA非常长。第二个问题涉及不同的分子如何在细胞内相互“交谈”，以产生控制细胞行为的信号。因此，我们将通过将信号分子家族的所有成员以不同的模式连接到DNA折纸瓦片上来解决这个问题，以显示一个成员与另一个成员的接近程度可能会产生影响。

项目成果

RSOB-16-0248.R1 - Supplementary text and figures from Coordinated regulation of the ESCRT-III component CHMP4C by the chromosomal passenger complex and centralspindlin during cytokinesis

RSOB-16-0248.R1 - 胞质分裂过程中染色体过客复合物和中枢轴蛋白对 ESCRT-III 成分 CHMP4C 的协调调节的补充文本和图片

• DOI: 10.6084/m9.figshare.4038198
• 发表时间: 2016
• 作者: Capalbo L

Coordinated regulation of the ESCRT-III component CHMP4C by the chromosomal passenger complex and centralspindlin during cytokinesis.

• DOI: 10.1098/rsob.160248
• 发表时间: 2016-10
• 期刊: Open biology
• 影响因子: 5.8
• 作者: Capalbo L;Mela I;Abad MA;Jeyaprakash AA;Edwardson JM;D'Avino PP
• 通讯作者: D'Avino PP

Functional analysis of the interface between the tandem C2 domains of synaptotagmin-1.

• DOI: 10.1091/mbc.e15-07-0503
• 发表时间: 2016-03-15
• 期刊: Molecular biology of the cell
• 影响因子: 3.3
• 作者: Evans CS;He Z;Bai H;Lou X;Jeggle P;Sutton RB;Edwardson JM;Chapman ER
• 通讯作者: Chapman ER

Purification of Recombinant ESCRT-III Proteins and Their Use in Atomic Force Microscopy and In Vitro Binding and Phosphorylation Assays.

重组 ESCRT-III 蛋白的纯化及其在原子力显微镜以及体外结合和磷酸化测定中的应用。

• DOI: 10.1007/978-1-4939-9492-2_15
• 发表时间: 2019
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Capalbo L

Coordinated regulation of the ESCRT-III component CHMP4C by the chromosomal passenger complex and centralspindlin during cytokinesis

胞质分裂过程中染色体过客复合物和中枢纺锤体对 ESCRT-III 成分 CHMP4C 的协调调节

• DOI: 10.17863/cam.6710
• 发表时间: 2016
• 作者: Capalbo L